Flexirubins are the unique type of bacterial pigments produced by the bacteria from the genus Chryseobacterium, which are used in the treatment of chronic skin disease, eczema etc. and may serve as a chemotaxonomic marker. Chryseobacterium artocarpi CECT 8497, an yellowish-orange pigment producing strain was investigated for maximum production of pigment by optimizing medium composition employing response surface methodology (RSM). Culture conditions affecting pigment production were optimized statistically in shake flask experiments. Lactose, l-tryptophan and KH2PO4 were the most significant variables affecting pigment production. Box Behnken design (BBD) and RSM analysis were adopted to investigate the interactions between variables and determine the optimal values for maximum pigment production. Evaluation of the experimental results signified that the optimum conditions for maximum production of pigment (521.64 mg/L) in 50 L bioreactor were lactose 11.25 g/L, l-tryptophan 6 g/L and KH2PO4 650 ppm. Production under optimized conditions increased to 7.23 fold comparing to its production prior to optimization. Results of this study showed that statistical optimization of medium composition and their interaction effects enable short listing of the significant factors influencing maximum pigment production from Chryseobacterium artocarpi CECT 8497. In addition, this is the first report optimizing the process parameters for flexirubin type pigment production from Chryseobacterium artocarpi CECT 8497.
A rotatable central composite design (CCD) was used to study the effect of cryoprotectants (skim milk, sucrose and lactose) on the survival rate of a probiotic Lactobacillus strain, L. reuteri C10, for poultry, during freeze-drying and storage. Using response surface methodology, a quadratic polynomial equation was obtained for response value by multiple regression analyses: Y = 8.59546-0.01038 X(1)-0.09382 X(2)-0.07771 X(3)-0.054861 X(1)(2)-0.04603 X(3)(2)-0.10938 X(1)X(2). Based on the model predicted, sucrose exerted the strongest effect on the survival rate. At various combinations of cryoprotectants, the viability loss of the cells after freeze-drying was reduced from 1.65 log colony forming units (CFU)/mL to 0.26-0.66 log CFU/mL. The estimated optimum combination for enhancing the survival rate of L. reuteri C10 was 19.5% skim milk, 1% sucrose and 9% lactose. Verification experiments confirmed the validity of the predicted model. The storage life of freeze-dried L. reuteri C10 was markedly improved when cryoprotectants were used. At optimum combination of the cryoprotectants, the survival rates of freeze-dried L. reuteri C10 stored at 4°C and 30°C for 6 months were 96.4% and 73.8%, respectively. Total viability loss of cells which were not protected by cryoprotectants occurred after 12 and 8 weeks of storage at 4°C and 30°C, respectively.
Changes in blood glucose are hypothesized to influence cognitive performance and these changes can be affected by certain nutrients. This double-blind 4-period cross-over study evaluated the effects of a slow-release modified sucrose (isomaltulose) in combination with a high concentration of lactose on cognitive performance of 5-6 year old children. Thirty children received a standard growing upmilk (Std GUM), reformulated growing up milk (Reform GUM), standard growing up milk with lactose-isomaltulose (Iso GUM), and a standard glucose drink (Glucose). The CDR System, a computerised cognitive assessment system, was used to assess various measures of attention and memory of the children at baseline (T=0), 60 (T=1), 120 (T=2), and 180 (T=3) minutes following the intake of test products. Overall, there was a decline in performance over the morning on almost every cognitive task. Children showed better attention following consumption of Iso GUM compared to Std GUM but attention was not significantly different than Reform GUM and glucose. Also, Iso GUM conferred a beneficial effect over both Reform GUM and glucose on sensitivity index of numeric working memory with no difference observed between Iso GUM and Std GUM. Surprisingly, glucose group showed lowest decline in the sensitivity index of spatial working memory and highest speed in picture recognition, although the latter was significantly better than Reform GUM only. For speed of spatial working memory, Reform GUM had the lowest decline but was significantly different only with Std GUM. There was, however, no significant difference among conditions for continuity of attention, speed of numeric working memory and picture recognition sensitivity. Despite the small sample size, the findings are intriguing as carbohydrate composition seems to influence some aspects of cognitive performance such as attention and memory. However, further studies are needed to confirm these findings.
The clinical response and the histological changes in the mucosa of the small bowel in response to continued feeding with cows' milk protein were assessed over a period of 2-6 weeks in 24 infants who had shown histological changes without immediate clinical symptoms after challenge with a diet containing cows' milk protein. Twenty of the 24 infants (83%) thrived well on cows' milk protein. Jejunal biopsy specimens taken six to eight weeks after the initial biopsy showed histological improvement in all 20 infants compared with biopsy specimens taken soon after the challenge, which had shown mucosal damage. The mucosa had returned to normal in 12, was mildly abnormal in seven, and moderately abnormal in one. Corresponding improvements in the activities of mucosal enzymes were seen. In four of the 24 infants (17%) symptoms developed between three and six weeks. Histological examination of the jejunal biopsy specimens showed that mucosal damage had progressed in two, and remained the same in two; moreover, the disaccharidase activities remained depressed. The present study shows that most infants with enteropathy caused by sensitivity to cows' milk protein but without clinical symptoms develop tolerance to the protein and the mucosa returns to normal despite continued feeding with cows' milk protein.
1,256 malnourished children, aged seven to ten years, were selected for study, 575 from Kuala Langat, 416 from Wilayah Persekutuan and 265 from Ulu Selangor. Ninety-three percent of the children were from low socioeconomic groups with large family size, and most of their parents had only primary or no formal education. During the study period, children in Kuala Langat received daily milk supplementation for five days per week, those in Wilayah Persekutuan for two days per week, while those in Ulu Selangor did not receive any milk supplementation. The study shows that a majority of the malnourished primary school children liked to drink milk and that milk intolerance was not a problem among them.
Chitosan nanoparticles are exhalation prone and agglomerative to pulmonary inhalation. Blending nanoparticles with lactose microparticles (∼5 µm) could mutually reduce their agglomeration through surface adsorption phenomenon. The chitosan nanoparticles of varying size, size distribution, zeta potential, crystallinity, shape and surface roughness were prepared by spray drying technique as a function of chitosan, surfactant and processing conditions. Lactose-polyethylene glycol 3000 (PEG3000) microparticles were similarly prepared. The chitosan nanoparticles, physically blended with fine lactose-PEG3000 microparticles, exhibited a comparable inhalation performance with the commercial dry powder inhaler products (fine particle fraction between 20% and 30%). Cascade impactor analysis indicated that the aerosolization and inhalation performance of chitosan nanoparticles was promoted by their higher zeta potential and circularity, and larger size attributes of which led to reduced inter-nanoparticulate aggregation and favored nanoparticles interacting with lactose-PEG3000 micropaticles that aided their delivery into deep and peripheral lungs.
The utilization of natural mica as a biocatalyst support in kinetic investigations is first described in this study. The formation of lactose caprate from lactose sugar and capric acid, using free lipase (free-CRL) and lipase immobilized on nanoporous mica (NER-CRL) as a biocatalyst, was evaluated through a kinetic study. The apparent kinetic parameters, K(m) and V(max), were determined by means of the Michaelis-Menten kinetic model. The Ping-Pong Bi-Bi mechanism with single substrate inhibition was adopted as it best explains the experimental findings. The kinetic results show lower K(m) values with NER-CRL than with free-CRL, indicating the higher affinity of NER-CRL towards both substrates at the maximum reaction velocity (V(max,app)>V(max)). The kinetic parameters deduced from this model were used to simulate reaction rate data which were in close agreement with the experimental values.
Chitin, as the main component of the exoskeleton of Arthropoda, is a highly available natural polymer that can be processed into various value-added products. Its most important derivative, i.e., chitosan, comprising β-1,4-linked 2-amino-2-deoxy-β-d-glucose (deacetylated d-glucosamine) and N-acetyl-d-glucosamine units, can be prepared via alkaline deacetylation process. Chitosan has been used as a biodegradable, biocompatible, non-antigenic, and nontoxic polymer in some in-vitro applications, but the recently found potentials of chitosan for in-vivo applications based on its biological activities, especially antimicrobial, antioxidant, and anticancer activities, have upgraded the chitosan roles in biomaterials. Chitosan approval, generally recognized as a safe compound by the United States Food and Drug Administration, has attracted much attention toward its possible applications in diverse fields, especially biomedicine and agriculture. Despite some favorable characteristics, the chitosan's structure should be customized for advanced applications, especially due to its drawbacks, such as low drug-load capacity, low solubility, high viscosity, lack of elastic properties, and pH sensitivity. In this context, derivatization with relatively inexpensive and highly available mono- and di-saccharides to soluble branched chitosan has been considered a "game changer". This review critically scrutinizes the emerging technologies based on the synthesis and application of lactose- and galactose-modified chitosan as two important chitosan derivatives. Some characteristics of chitosan derivatives and biological activities have been detailed first to understand the value of these natural polymers. Second, the saccharide modification of chitosan has been discussed briefly. Finally, the applications of lactose- and galactose-modified chitosan have been scrutinized and compared to native chitosan to provide an insight into the current state-of-the research for stimulating new ideas with the potential of filling research gaps.
In the attempt to create a delivery system for an alkali-cation stimulated drug release, a computational study was conducted, aiming for the evaluation of synthetic access towards glycolipid crown ethers analogs and their potential for coordination-induced changes of packing constraints for molecular assemblies. The results disfavor amide-linkages for the creation of macrocycles around the inter-glycosidic bond of a disaccharide. Conformational changes upon cation coordination of the macrocycle decrease the intersection area for easily accessible macrocycles based on lactose. This leads to shrinking intersection areas upon alkali complexation. Maltose-based analogs, on the other hand, exhibited the targeted increase of the glycolipid intersection area and, hence, may be considered as a promising resource.
Percolation theory has been used with great interest in understanding the design and characterization of dosage forms. In this study, work has been carried out to investigate the behavior of binary mixture tablets containing excipients of similar and different deformation properties. The binary mixture tablets were prepared by direct compression using lactose, polyvinyl chloride (PVC), Eudragit RS 100, and microcrystalline cellulose (MCC). The application of percolation theory on the relationships between compactibility, Pmax, or compression susceptibility (compressibility), gamma, and mixture compositions reveals the presence of percolation thresholds even for mixtures of similar deformation properties. The results showed that all mixture compositions exhibited at least one discreet change in the slope, which was referred to as the percolation threshold. The PVC/Eudragit RS100 mixture compositions showed significant percolation threshold at 80% (w/w) PVC loading. Two percolation thresholds were observed from a series of binary mixtures containing similar plastic deformation materials (PVC/MCC). The percolation thresholds were determined at 20% (w/w) and 80% (w/w) PVC loading. These are areas where one of the components percolates throughout the system and the properties of the tablets are expected to experience a sudden change. Experimental results, however, showed that total disruption of the tablet physical properties at the specified percolation thresholds can be observed for PVC/lactose mixtures at 20-30% (w/w) loading while only minor changes in the tablets' strength for PVC/MCC or PVC/Eudragit RS 100 mixtures were observed.
Controlled-release grade hydroxypropylmethylcellulose (HPMC) or xanthan gum (XG) and microcrystalline cellulose (MCC) were employed to prepare controlled-release diltiazem hydrochloride tablets. The similarity factor f2 was used for dissolution profile comparison using Herbesser 90 SR as a reference product. Drug release could be sustained in a predictable manner by modifying the content of HPMC or XG. Moreover, the drug release profiles of tablets prepared using these matrix materials were not affected by pH and agitation rate. The f2 values showed that only one batch of tablets (of diltiazem HCl, HPMC or XG, and MCC in proportions of 3.0:3.0:4.0) was considered similar to that of the reference product, with values above 50. The unbiased similarity factor f2* values were not much different from the f2 values, ascribing to a small dissolution variance of the test and reference products. The amount of HPMC or XG incorporated to produce tablets with the desired dissolution profile could be determined from the curves of f2 versus polymer content. Hence, the f2 values can be applied as screening and optimization tools during development of controlled-release preparations.
Functional nanomaterials have been pursued to assemble nanobiocatalysts since they can provide unique hierarchical nanostructures and localized nanoenvironments for enhancing enzyme specificity, stability and selectivity. Functionalized dendrimer-like hierarchically porous silica nanoparticles (HPSNs) was fabricated for assembling β-galactosidase nanobiocatalysts for bioconversion of lactose to galacto-oligosaccharides (GOS). The nanocarrier was functionalized with amino (NH2) and carboxyl (COOH) groups to facilitate enzyme binding, benchmarking with non-functionalized HPSNs. Successful conjugation of the functional groups was confirmed by FTIR, TGA and zeta potential analysis. HPSNs-NH2 showed 1.8-fold and 1.1-fold higher β-galactosidase adsorption than HPSNs-COOH and HPSNs carriers, respectively, with the highest enzyme adsorption capacity of 328mg/g nanocarrier at an initial enzyme concentration of 8mg/ml. The HPSNs-NH2 and β-galactosidase assembly (HPSNs-NH2-Gal) demonstrated to maintain the highest activity at all tested enzyme concentrations and exhibited activity up to 10 continuous cycles. Importantly, HPSNs-NH2-Gal was simply recycled through centrifugation, overcoming the challenging problems of separating the nanocarrier from the reaction medium. HPSNs-NH2-Gal had distinguished catalytic reaction profiles by favoring transgalactosylation, enhancing GOS production of up to 122g/l in comparison with 56g/l by free β-galactosidase. Furthermore, it generated up to 46g/l GOS at a lower initial lactose concentration while the free counterpart had negligible GOS production as hydrolysis was overwhelmingly dominant in the reaction system. Our research findings show the amino-functionalized HPSNs can selectively promote the enzyme activity of β-galactosidase for transgalactosylation, which is beneficial for GOS production.
Lactic acid bacteria is well known for it uses as starter culture in various fermented food, and it functions as a good natural antimicrobial agent. Cincaluk, a Malaysian fermented shrimp product commonly found in traditional dishes is commonly enriched with LAB. Out of 50 colonies from a local cincaluk, 7 strains were successfully isolated and shown to be positive in lactose utilization and catalase tests. The majority of the isolates from cincaluk showed Gram-positive cocci morphology and belonged to the group Staphyloccoccus spp. By using agar disc diffusion method, the anti-bacterial properties of these isolates (namely isolate 1, 2, 3, 4, 5, 6, and 7) moderately inhibited the growth of several pathogenic strains, i.e., Escherichia coli, Staphylococcus aureus, Salmonella typhimurium and Bacillus subtilis which were used as indicator bacteria. Other than isolates 1, 2, 3 and 5; the 16S rRNA gene for isolate 6 and 7 were successfully amplified. The 16S rRNA gene fragment from isolate 7 was successfully cloned and sequenced. Based on rRNA sequences, both isolates 6 and 7 belonged to the group Staphylococcus piscifermentans, a rare strain previously reported to be specifically isolated exclusive from fish sources.
The aim of this study was to determine the specific content and type of sugars in selected commercial and traditional kuih in Klang Valley. The selection of the kuih was based on the validated Food Frequency Questionnaire (FFQ) for sugar. The selected commercial kuih was doughnut coated with sugar (Big Apple) while the ten traditional kuih samples consisted of kuih bingka ubi, kuih kasui, kuih keria, kuih koci, kuih lapis, kuih lopes, kuih onde-onde, kuih sagu, kuih seri muka and kuih talam. The doughnut coated with sugar (Big Apple) was purchased from Big Apple Donuts and Coffee franchise at two different locations, while the traditional kuih were randomly bought from stalls, cafeterias and restaurants around Kuala Lumpur and Rawang. The types and amount of sugar were determined using High Performance High Chromatography (HPLC) with a refractive index (RI) detector. Results showed that doughnut coated with sugar (Big Apple) has the highest starch content (22.6±0.3 g/100g) and kuih keria contained the highest available carbohydrate (41.5±1.7 g/100g), comprising of 24.2±2.4 g/100g total sugar and 17.3±0.7 g/100g of starch. The least available carbohydrate content was found in kuih talam (20.0±0.5 g/100g), which was 50% lower than the one in kuih keria. Major individual sugars detected in all kuih samples were consisted of sucrose (60.0%), glucose (16.2%), fructose (14.0%), maltose (9.5%) and lactose (0.3%). Majority of the kuih samples (90.9%) in this study can be categorized as medium sugar while only kuih keria was categorized as high sugar. Based on the two main ingredients (sugar and flour) used in the preparation of kuih, results showed that all kuih samples can be categorized as medium sugarmedium starch. In conclusion, this study served as a guideline by locals in selecting kuih of
different sugar levels.
This review explores the digestibility of lactose by Malaysians, and the value of milk and other milk-derived products as sources of appropriate nutrition for Malaysians. Increased calcium intake through consumption of milk is an effective mechanism for increasing calcium uptake from the diet and thereby minimising the risk of development of osteoporosis in later life. Detailed information about rates of lactose intolerance, and adaptation to dietary lactose and its consequences for Malaysians, will help in the formulation of dietary advice, and improve commerial food manufaturing practice and Government policy
directed to the minimization of rates of osteoporosis, which presents a substantial morbidity risk to elderly female Asians in particular.
Conventional melt pelletization and granulation processes produce round and dense, and irregularly shaped but porous agglomerates respectively. This study aimed to design centrifugal air-assisted melt agglomeration technology for manufacture of spherical and yet porous "granulets" for ease of downstream manufacturing and enhancing drug release. A bladeless agglomerator, which utilized shear-free air stream to mass the powder mixture of lactose filler, polyethylene glycol binder and poorly water-soluble tolbutamide drug into "granulets", was developed. The inclination angle and number of vane, air-impermeable surface area of air guide, processing temperature, binder content and molecular weight were investigated with reference to "granulet" size, shape, texture and drug release properties. Unlike fluid-bed melt agglomeration with vertical processing air flow, the air stream in the present technology moved centrifugally to roll the processing mass into spherical but porous "granulets" with a drug release propensity higher than physical powder mixture, unprocessed drug and dense pellets prepared using high shear mixer. The fast-release attribute of "granulets" was ascribed to porous matrix formed with a high level of polyethylene glycol as solubilizer. The agglomeration and drug release outcomes of centrifugal air-assisted technology are unmet by the existing high shear and fluid-bed melt agglomeration techniques.
The conventional powder flow testers require sample volumes larger than 40g and are met with experimental hiccups due to powder cohesion. This study designed a gas-pressurized dispersive powder flow tester where a high velocity air is used to disaggregate powder (9g) and eliminate its cohesion. The pressurized gas entrained solid particles leaving an orifice where the distance, surface area, width and weight of particle dispersion thereafter are determined as flow index. The flow indices of seven lactose grades with varying size, size distribution, shape, morphology, bulk and tapped densities characteristics were examined. They were compared against Hausner ratio and Carr's index parameters of the same powder mass. Both distance and surface area attributes of particle dispersion had significant negative correlations with Hausner ratio and Carr's index values of lactose. The distance, surface area and ease of particle dispersion varied proportionately with circular equivalent, surface weighted mean and volume weighted mean diameters of lactose, and inversely related to their specific surface area and elongation characteristics. Unlike insensitive Hausner ratio and Carr's index, an increase in elongation property of lactose particles was detectable through reduced powder weight loss from gas-pressurized dispersion as a result of susceptible particle blockage at orifice. The gas-pressurized dispersive tester is a useful alternative flowability measurement device for low volume and cohesive powder.
Controlled release buccal patches were fabricated using Eudragit NE40D and studied. Various bioadhesive polymers, namely hydroxypropylmethyl cellulose, sodium carboxymethyl cellulose and Carbopol of different grades, were incorporated into the patches, to modify their bioadhesive properties as well as the rate of drug release, using metoprolol tartrate as the model drug. The in-vitro drug release was determined using the USP 23 dissolution test apparatus 5 with slight modification, while the bioadhesive properties were evaluated using texture analyzer equipment with chicken pouch as the model tissue. The incorporation of hydrophilic polymers was found to affect the drug release as well as enhance the bioadhesiveness. Although high viscosity polymers can enhance the bioadhesiveness of the patches, they also tend to cause non-homogeneous distribution of the polymers and drug, resulting in non-predictable drug-release rates. Of the various bioadhesive polymers studied, Cekol 700 appeared to be most satisfactory in terms of modifying the drug release and enhancement of the bioadhesive properties.
This study developed a tester where the powder flow was characterized using a low sample mass (2 g) and impact instead of dispersion mechanism to mitigate test space constraint. An impact chamber was established where the test powder bed of seven lactose grades was weight-impacted to produce impact crater and ejecta, and imaged quantitatively to determine crater profiling signature (crater depth), regional topography (ejecta roughness), Otsu threshold (bed continuity) and edge segmentation (bed deformation). The Hausner ratio (HR) and Carr's index (CI) values of lactose, and their powder dispersion distance and surface area characteristics evaluated by gas-pressurized dispersibility test were examined as reference method. The crater signature profiling and regional topography were correlated to HR, CI, dispersive distance and surface area. A poorer powder flow was characterized by higher values of crater signature profiling, regional topography, HR, CI, and lower dispersive distance and surface area. The crater signature profiling and regional topography values were higher with smaller and rougher lactose particles that were cohesive. The powder impact flow is a viable non-dispersive approach to characterize powder flowability using a small sample mass and test space.